Pooling of Nasopharyngeal Swab Samples To Overcome a Global Shortage of Real-Time Reverse Transcription-PCR COVID-19 Test Kits

More, Sunil; Narayanan, Sai; Patil, Girish; Ghosh, Parna; Pushparaj, Samuel; Cooper, Emily; Ritchey, Jerry; Cheruvu, Vinay K.; Kaul, Anil; Ramachandran, Akhilesh

The global outbreak and rapid spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) have created an urgent need for large-scale testing of populations. There is a demand for high-throughput testing protocols that can be used for efficient and rapid testing of clinical specimens. We evaluated a pooled PCR protocol for testing nasopharyngeal (NP) swabs using known positive/negative and untested clinical samples that were assigned to pools of 5 or 10. In total, 630 samples were used in this study. Individual positive samples with cycle threshold (C(T)) values as high as 33 could be consistently detected when pooled with 4 negative samples (pool of 5), and individual positive samples with C(T) values up to 31 could be consistently detected when pooled with 9 negative samples (pool of 10). Pooling of up to 5 samples can be employed in laboratories for the diagnosis of COVID-19 for efficient utilization of resources, rapid screening of a greater number of people, and faster reporting of test results.